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Method for identifying drought resistance of highland barley by utilizing semi-quantitative PCR method of HVA1 gene

A drought resistance, semi-quantitative technology, applied in the field of identification of drought resistance of highland barley, can solve the problems of plant drought resistance without physiological and biochemical indicators, cumbersome methods, and difficult identification of plant drought resistance

Inactive Publication Date: 2013-07-03
QINGHAI UNIVERSITY
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Problems solved by technology

[0003] At present, the identification of drought resistance mostly adopts physiological and biochemical methods, which are cumbersome, and it is difficult to identify the drought resistance of plants only by one or two physiological indicators, so it takes a long time, and there is no fixed physiological and biochemical indicators to determine the drought resistance of plants sex

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  • Method for identifying drought resistance of highland barley by utilizing semi-quantitative PCR method of HVA1 gene
  • Method for identifying drought resistance of highland barley by utilizing semi-quantitative PCR method of HVA1 gene
  • Method for identifying drought resistance of highland barley by utilizing semi-quantitative PCR method of HVA1 gene

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Embodiment Construction

[0017] The present invention will be described in detail below in conjunction with specific embodiments.

[0018] Table 1 Primer name and sequence

[0019]

[0020] Primer P1 was designed according to the sequence of barley HVA1 gene (X78205) published by GenBank, and a pair of primers P2 were designed for barley actin (β-Actin) gene (ID: AY145541) using Primer5.0. synthesis.

[0021] Using drought-resistant purple highland barley and drought-sensitive hemp highland barley as materials, select highland barley seeds with consistent plumpness and no damage from diseases and insect pests (two varieties with strong drought resistance and weak drought resistance), soak in clean water for 1 hour, and use 0.5% high Potassium manganate was sterilized for 30 minutes, washed with water, and the seeds were immersed in distilled water and placed in a refrigerator at 4°C for 24 hours. Take 20 seeds each and sow them in a petri dish with a diameter of 10 cm, covered with 3 layers of di...

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Abstract

The invention discloses a method for identifying the drought resistance of highland barley by utilizing a semi-quantitative PCR (Polymerase Chain Reaction) method of an HVA1 gene, which comprises the following steps: (1) carrying out stress treatment for 4 days by utilizing 0-30 percent of PEG (Polyethyleneglycol) on the two-leaf stage of the highland barley; (2) carrying out total RNA (Ribose Nucleic Acid) extraction and reverse transcription on a blade by adopting a total RNA extraction reagent kit and a reverse transcription reagent kit from Shanghai Sangon; (3) amplifying a target gene HVA1 through taking a barley actin (Beta-Actin) gene as a reference gene; (4) carrying out quadratic function fitting on a relative expression map of the obtained (Beta-Actin) gene by adopting Gel-pro software; and (5) calculating the highest relative expression quantity of the HVA1 gene or the corresponding PEG concentration according to a fitting curve, and if the numerical value is larger, a highland barley variety with strong drought resistance is determined.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for identifying the drought resistance of highland barley by using the semi-quantitative PCR method of the HVA1 gene. Background technique [0002] Late embryogenesis abundant proteins (LEA proteins) in higher plants are a class of small molecular specific polypeptides produced in the later stages of seed development. These proteins are closely related to plant dehydration tolerance, and are affected by plant developmental stages, ABA and dehydration. Regulation of signaling (Sun et al., 2003; Ramanjulu, 2002). Since 1981, a group of abundantly expressed mRNA was isolated from the cotton cotyledon in the later stage of embryonic development for the first time, named leamRNA, and its expression product, LEA protein (Dure et al., 1981), has been isolated from various Many new LEA proteins have been discovered in plants. According to the homology of amino acid sequences and s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 姚晓华吴昆仑蒋礼玲王显萍任又成
Owner QINGHAI UNIVERSITY
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